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Rnai-based functional analysis of bursicon genes related to cuticle pigmentation in a ladybird beetle 基于 Rnai 的瓢虫角质层色素相关囊状基因功能分析
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-08-22 DOI: 10.1016/j.jinsphys.2024.104696
Lan-Lan Liao, Wen-Ze Li, Lin Jin, Guo-Qing Li

In arthropods, the binding of a bursicon (encoded by burs and pburs) heterodimer or homodimer to a leucine-rich repeat-containing G protein coupled receptor LGR2 (encoded by rk) can activate many physiological processes, especially cuticle pigmentation during insect ecdysis. In the current paper, we intended to ascertain whether bursicon signaling mediates body coloration in the 28-spotted larger potato ladybird, Henosepilachna vigintioctomaculata, and if so, by which way bursicon signal governs the pigmentation. The high expression of Hvburs, Hvpburs and Hvrk occurred in the young larvae, pupae and adults, especially in the head and ventral nerve cord. RNA interference (RNAi) aided knockdown of Hvburs, Hvpburs or Hvrk in the prepupae caused similar phenotypic defects. The pigmentation of the resultant adults was affected, with significantly reduced dark areas on the sternums. Moreover, the accumulated mRNA levels of two sclerotin biosynthesis genes, aspartate 1-decarboxylase gene Hvadc and N-β-alanyldopamine synthase gene Hvebony, were significantly increased in the Hvburs, Hvpburs or Hvrk RNAi beetles. Furthermore, depletion of either Hvadc or Hvebony could completely rescue the impaired coloration on the sternums of Hvpburs RNAi adult. Our results supported that bursicon heterodimer-mediated signal regulate cuticle pigmentation. The bursicon signaling may tune the ratio of melanins (dark/black, brown) to sclerotins (light yellow, colorless) exerting its regulative role in the pigmentation of H. vigintioctomaculata sternums.

在节肢动物中,法氏囊异源二聚体(由法氏囊和法氏囊编码)或同源二聚体与含亮氨酸丰富重复的 G 蛋白偶联受体 LGR2(由 rk 编码)的结合可激活许多生理过程,尤其是昆虫蜕皮过程中的角质层色素沉着。在本论文中,我们试图弄清法氏囊信号是否介导了 28 点大马铃薯瓢虫(Henosepilachna vigintioctomaculata)的身体着色,如果是,法氏囊信号是通过何种方式控制着色的。Hvburs、Hvpburs和Hvrk在幼虫、蛹和成虫中均有高表达,尤其是在头部和腹侧神经索。在RNAi干扰(RNAi)的帮助下敲除幼虫体内的Hvburs、Hvpburs或Hvrk会导致类似的表型缺陷。成虫的色素沉着受到影响,胸骨上的暗色区域明显减少。此外,在 Hvburs、Hvpburs 或 Hvrk RNAi 甲虫中,两个硬脂素生物合成基因(天冬氨酸 1-脱羧酶基因 Hvadc 和 N-β-丙氨酰多巴胺合成酶基因 Hvebony)的累积 mRNA 水平显著增加。此外,消耗 Hvadc 或 Hvebony 基因可以完全修复 Hvpburs RNAi 成虫胸骨上受损的着色。我们的研究结果表明,囊状体异源二聚体介导的信号调节角质层色素沉着。Bursicon 信号可能会调节黑色素(深/黑,棕色)和硬蛋白(浅黄色,无色)的比例,从而在 H. vigintioctomaculata 胸骨色素沉着中发挥调节作用。
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引用次数: 0
Microplastic exposure reduced the defecation rate, altered digestive enzyme activities, and caused histological and ultracellular changes in the midgut tissues of the ground beetle (Blaps polychresta) 接触微塑料降低了地鳖虫(Blaps polychresta)的排便率,改变了消化酶活性,并导致中肠组织发生组织学和超细胞变化。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-08-17 DOI: 10.1016/j.jinsphys.2024.104697
Samar El kholy , Taiwo Ayorinde , Christie M. Sayes , Yahya Al Naggar

Concerns about microplastic (MP) pollution in terrestrial systems are increasing. It is believed that the overall amount of MPs in the terrestrial system could be 4–23 times higher than that in the ocean. Agricultural ecosystems are among the most polluted areas with MPs. Terrestrial organisms such as ground beetles, will be more vulnerable to MPs in various agricultural soil types because they are common in garden and agricultural areas. Therefore, this work aims to assess for the first time the potential adverse effects of chronic exposure for 30 days of ground beetles to a field-realistic concentration of 2 % (w/w) of three different irregularly shaped MPs polymers: Polystyrene (PS), polyethylene terephthalate (PET), and polyamide 6 (PA; i.e., nylon 6) on their health. The results showed no effect on beetle survival; nevertheless, there was a decrease in beetle defecation rate, particularly in beetles exposed to PS-MPs, and a change in the activity of midgut digestive enzymes. The effects on digestive enzymes (amylase, protease, lipase, and α-glucosidase) were polymer and enzyme specific. Furthermore, histological and cytological studies demonstrated the decomposition of the midgut peritrophic membrane, as well as abnormally shaped nuclei, vacuolation, disordered microvilli, necrosis of goblet and columnar cells, and necrosis of mitochondria in midgut cells. Given the importance of ground beetles as predators in most agricultural and garden settings, the reported adverse impacts of MPs on their health may impact their existence and ecological functions.

人们对陆地系统中微塑料(MP)污染的关注与日俱增。据信,陆地系统中的微塑料总量可能是海洋中的 4-23 倍。农业生态系统是受到微塑料污染最严重的地区之一。由于地鳖虫等陆生生物常见于花园和农业区,因此它们更容易受到各种农业土壤中 MPs 的影响。因此,这项工作旨在首次评估地鳖虫在长期接触 2%(重量/湿重)浓度的三种不同不规则形状的 MPs 聚合物 30 天后可能产生的不良影响:聚苯乙烯(PS)、聚对苯二甲酸乙二酯(PET)和聚酰胺 6(PA,即尼龙 6)对甲虫健康的潜在不利影响。结果表明,聚苯乙烯(PS)和聚对苯二甲酸乙二酯(PET)以及聚酰胺 6(PA,即尼龙 6)对甲虫的存活率没有影响;不过,甲虫的排便率有所下降,尤其是接触聚苯乙烯-MPs 的甲虫,中肠消化酶的活性也发生了变化。对消化酶(淀粉酶、蛋白酶、脂肪酶和 α-葡萄糖苷酶)的影响具有聚合物和酶的特异性。此外,组织学和细胞学研究表明,中肠周围膜分解,中肠细胞核形状异常、空泡化、微绒毛紊乱、鹅口疮细胞和柱状细胞坏死以及线粒体坏死。鉴于地鳖虫在大多数农业和园林环境中作为天敌的重要性,报告的 MPs 对其健康的不利影响可能会影响其生存和生态功能。
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引用次数: 0
Utilization of a Clark electrode device as a respirometer for small insects: A convincing test on ants allowing to detect discontinuous gas exchange 利用克拉克电极装置作为小型昆虫的呼吸计:在蚂蚁身上进行的令人信服的测试,可以检测到不连续的呼吸。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-08-17 DOI: 10.1016/j.jinsphys.2024.104698
Maïly Kervella , Céline Cansell , François Criscuolo , Frederic Bouillaud

Respirometry provides a direct measure of an organism’s O2 consumption rate (VO2), which is a significant component of its metabolic rate (energy expenditure). Amongst ants, variations in lifespan between different social castes (such as workers and queens) can be substantial, varying depending on the species. As metabolic rate is higher in short-living species, we aimed to determine how VO2 and longevity may have coevolved within ant casts. Measuring VO2 in such tiny animal models can be challenging, and as a first methodological step, we validate the use of a Clark electrode, initially designed for measuring mitochondrial respiration control pathways, for assessing VO2 in ants within a sealed chamber. This was done by comparing it with stop-flow VO2 and CO2 production, using a traditional indirect calorimetry device. The global aim is to provide a reliable protocol to conduct accurate comparisons of metabolic rates within and among ant species. As expected, using the Clark electrode entails high time resolution and revealed that queens and workers exhibited discontinuous gas exchange, with episodes of apnea lasting up to 20 min.

呼吸测定法可直接测量生物体的氧气消耗率(VO2),这是生物体代谢率(能量消耗)的重要组成部分。在蚂蚁中,不同社会等级(如工蚁和蚁后)之间的寿命差异很大,因物种而异。由于短寿命物种的新陈代谢率较高,我们的目标是确定 VO2 和寿命在蚂蚁种群中是如何共同进化的。在如此微小的动物模型中测量 VO2 可能具有挑战性,作为方法论的第一步,我们验证了使用克拉克电极(最初设计用于测量线粒体呼吸控制途径)评估密封舱中蚂蚁的 VO2。为此,我们使用传统的间接量热装置,将其与停止流动的 VO2 和二氧化碳产生量进行了比较。总体目标是提供一种可靠的方案,对蚂蚁物种内部和物种之间的代谢率进行准确的比较。不出所料,使用克拉克电极可以获得很高的时间分辨率,并发现蚁后和工蚁表现出不连续的呼吸,呼吸暂停可持续长达 20 分钟。
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引用次数: 0
Differential expression of fibroin-related genes in middle silk glands is induced by dietary differences in a strain-dependent manner in Bombyx mori 中丝腺体中纤维蛋白相关基因的差异表达是由膳食差异诱导的,而膳食差异又是以品系为基础的。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-08-16 DOI: 10.1016/j.jinsphys.2024.104695
Tsuneyuki Tatsuke, Shuichiro Tomita

The silkworm (Bombyx mori) is a model organism for lepidopteran insects. It is an oligophagous insect that primarily feeds on mulberry leaves and has industrial use for the production of raw silk. The development of artificial diets has provided an alternative nutrient source for silkworms; however, one significant issue is that the production of cocoons is lower in silkworms reared on artificial diets compared with those reared on mulberry leaves. The differences in the silk gland in the late-stage fifth instar silkworm larvae, when silk synthesis is most active, between those raised on artificial diets and mulberry leaves, are unknown. In this study, we identified differences in the transcriptomes of the middle and posterior silk glands of fifth instar day five silkworm larvae reared on artificial diets compared with those reared on mulberry leaves using three strains: Daizo, Nichi01, and J137 × C146. We found that the silk-related genes fibrohexamerin (fhx), fibroin-light-chain (fibL), and fibroin-heavy-chain (fibH) in the middle silk gland, and ser1 in the posterior silk gland, were differentially expressed in a strain-dependent manner. In silkworms reared on artificial diets, fhx, fibL, and fibH in the middle silk gland were upregulated in Nichi01 and downregulated in J137 × C146, whereas ser1 in the posterior silk gland was upregulated in J137 × C146 compared with silkworms reared on mulberry leaves. Our results demonstrate that the diet and strain of silkworm larvae affect the expression of genes related to silk production in their silk glands during the late fifth instar stage.

家蚕(Bombyx mori)是鳞翅目昆虫的模式生物。它是一种寡食性昆虫,主要以桑叶为食,可用于工业生产生丝。人工饲料的开发为蚕提供了另一种营养来源;然而,一个重要的问题是,与桑叶饲养的蚕相比,人工饲料饲养的蚕产茧量较低。人工饲料和桑叶饲养的蚕在第五龄幼虫后期(蚕丝合成最活跃的时期)的丝腺差异尚不清楚。在这项研究中,我们利用三个品系,确定了人工饲料饲养的第五龄第五天蚕幼虫与桑叶饲养的第五龄第五天蚕幼虫中后期丝腺转录组的差异:大藏、Nichi01 和 J137 × C146。我们发现,中丝腺中与丝有关的基因纤维六聚氨基甲酸(fhx)、纤维蛋白轻链(fibL)和纤维蛋白重链(fibH)以及后丝腺中的 ser1 都以依赖于品系的方式差异表达。与桑叶饲养的蚕相比,人工日粮饲养的蚕,中丝腺中的fhx、fibL和fibH在Nichi01中上调,在J137 × C146中下调,而后丝腺中的ser1在J137 × C146中上调。我们的研究结果表明,蚕幼虫的食性和品系会影响其第五龄后期丝腺中产丝相关基因的表达。
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引用次数: 0
The morphology of the pygidial glands and the chemical composition of their secretions of four sphodrine ground beetle species (Carabidae: Platyninae) 四种鞘翅目地鳖虫(Carabidae: Platyninae)的鳃腺形态及其分泌物的化学成分。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-08-03 DOI: 10.1016/j.jinsphys.2024.104685
Sofija Vranić , Ljubodrag Vujisić , Nikola Vesović , Marina Todosijević , Dejan Pantelić , Danica Pavlović , Stefan Ivanović , Marija Vasović , Srećko Ćurčić

Ground beetles possess a pair of pygidial glands that produce and release secretions that play an important role in defense against predators. The morphology of these glands and the chemical composition of their products were studied in four species of the tribe Sphodrini: Calathus (Calathus) fuscipes (Goeze, 1777), C. (Neocalathus) cinctus Motschulsky, 1850, C. (N.) melanocephalus (Linnaeus, 1758) and Laemostenus (Antisphodrus) elongatus (Dejean, 1828). The morphological analyzes of the glands of the four taxa mentioned were carried out for the first time using bright-field and nonlinear microscopy. All morphological structures were precisely measured and photographed. The pygidial gland secretions of C. (C.) fuscipes and L. (A.) elongatus were analyzed for the first time using gas chromatography-mass spectrometry. A total of 30 compounds were detected from the extracts of pygidial gland secretions of the four Sphodrini species studied. The simplest chemical mixture was found in L. (A.) elongatus, while the most complex secretion was that of C. (C.) fuscipes. 1-Undecanol, which we were able to detect in all taxa examined here, and dodecyl butyrate, which was detected in the three Calathus species, have never before been detected in the secretions of ground beetles.

地鳖拥有一对鳃腺,可产生和释放分泌物,在抵御天敌方面发挥着重要作用。我们对 Sphodrini 科的四个物种的这些腺体的形态及其产物的化学成分进行了研究:Calathus (Calathus) fuscipes (Goeze, 1777)、C. (Neocalathus) cinctus Motschulsky, 1850、C. (N.) melanocephalus (Linnaeus, 1758) 和 Laemostenus (Antisphodrus) elongatus (Dejean, 1828)。首次使用明视野和非线性显微镜对上述四个类群的腺体进行了形态分析。所有形态结构都经过精确测量和拍照。首次使用气相色谱-质谱法分析了 C. (C. fuscipes) 和 L. (A.) elongatus 的鳃腺分泌物。从所研究的四个 Sphodrini 种类的鳃腺分泌物提取物中共检测到 30 种化合物。L. (A.) elongatus 的化学混合物最为简单,而 C. (C.) fuscipes 的分泌物最为复杂。1- 十一烷醇和十二烷基丁酸酯以前从未在地鳖虫的分泌物中检测到过。
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引用次数: 0
Effects of parental diet on Mormon cricket egg diapause, embryonic development rate, and periodic outbreaks 父母饮食对蟋蟀卵休眠、胚胎发育率和周期性爆发的影响
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-28 DOI: 10.1016/j.jinsphys.2024.104681
Robert B. Srygley
<div><p>Transgenerational phenotypic modification can alter organismal fitness, population demographics, and community interactions. For ectotherms, both dietary composition and temperature have important effects on organismal fitness, but they are rarely investigated together. Mormon crickets <em>Anabrus simplex</em> are capable of diapausing as eggs in the soil for multiple years with duration largely dependent on cumulative heat units or degree days. Because Mormon crickets can be abundant in the landscape in one year and disappear suddenly the next, I asked: does parental nutrition affect the duration of egg diapause? Beginning in the ultimate nymphal instar, Mormon crickets were fed a diet high in protein, one equal in protein to carbohydrate, or a diet high in carbohydrates and the time for eggs to develop after they were laid was measured. If parental nutrition affects temperature-sensitive egg diapause, then that change in sensitivity to temperature might also alter the relationship between embryonic development rate and temperature. I asked: does parental nutrition affect embryonic development rate as a function of temperature? To this end, I manipulated densities of Mormon cricket nymphs and protein-rich prey (grasshoppers) in field cages, collected eggs from the adult Mormon crickets, and measured the optimal temperature, maximum development rate, and thermal breadth for embryonic development of the offspring. I found that Mormon crickets fed a high protein diet laid eggs with shorter diapause. Consistent with this long-term result, those housed with the most grasshoppers to eat laid eggs that had the fastest maximum development rate, whereas those without grasshoppers laid eggs with slower maximum developmental rates but the broadest thermal breadth. Eggs from Mormon crickets housed with intermediate levels of grasshopper densities had a decline in peak development rate with an increase in density. In addition, Mormon crickets housed with more conspecifics laid eggs with faster development rates, whereas thermal breadth and the temperature optima were not affected by cricket density. As predicted, Mormon cricket diets significantly affected egg diapause and development rates. Contrary to expectations based on observed changes in diet preferences during a Mormon cricket outbreak, Mormon crickets fed high protein diets laid eggs with significantly shorter egg diapause and significantly faster egg development rates. Interestingly, doubling of Mormon cricket density caused eggs to develop in nearly half the time. This latter result indicates that Mormon cricket aggregations promote rapid development of progeny. Moreover, the tight, linear structure of migratory bands in which females intermittently stop to lay eggs assures that the progeny hatch and develop in dense cohorts. In this manner, the banding behavior might carry-over into subsequent generations as long as cohorts are dense and protein is available. With band thinning or protein
跨代表型改变可改变生物体的适应性、种群人口统计学和群落相互作用。对于外温动物来说,食物组成和温度都会对生物的适应性产生重要影响,但很少同时对它们进行研究。摩门蟋 Anabrus simplex 能够以卵的形式在土壤中腹泻多年,腹泻持续时间主要取决于累积热量单位或度日。由于莫门蟋蟀可能在某一年大量繁殖,而第二年就会突然消失,因此我问:亲代营养是否会影响卵的休眠期?从蛹的末期开始,给摩门蟋喂高蛋白食物、蛋白质与碳水化合物含量相等的食物或高碳水化合物食物,并测量产卵后卵的发育时间。如果亲代营养会影响对温度敏感的卵停产,那么对温度敏感性的变化也可能会改变胚胎发育速度与温度之间的关系。我的问题是:亲代营养是否会影响胚胎发育率,使其成为温度的函数?为此,我操纵了田间笼子中的摩门蟋若虫和富含蛋白质的猎物(蚱蜢)的密度,收集了摩门蟋成虫的卵,并测量了后代胚胎发育的最佳温度、最大发育率和热广度。我发现,喂食高蛋白食物的摩门蟋产下的卵停滞期较短。与这一长期结果相一致的是,吃蚱蜢最多的蟋蟀产下的卵具有最快的最大发育速度,而不吃蚱蜢的蟋蟀产下的卵最大发育速度较慢,但热广度最大。饲养蚱蜢密度处于中等水平的摩门蟋产下的卵,随着密度的增加,最高发育率有所下降。此外,与更多同种蟋蟀一起饲养的摩门蟋产卵的发育速度更快,而热广度和最适温度则不受蟋蟀密度的影响。正如预测的那样,摩门蟋的饮食对卵的休眠和发育率有显著影响。与根据摩门蟋爆发期间观察到的食物偏好变化所做的预期相反,饲喂高蛋白食物的摩门蟋产卵的休眠期明显更短,卵的发育速度明显更快。有趣的是,摩门蟋密度增加一倍,卵的发育时间缩短近一半。后一项结果表明,摩门蟋的聚集促进了后代的快速发育。此外,雌性蟋蟀间歇性地停下来产卵,这种紧密的线性迁移带结构确保了后代在密集的群体中孵化和发育。通过这种方式,只要同群密度高且蛋白质充足,带状行为就有可能延续到后代。如果稀疏带状卵或限制蛋白质摄入,雌虫就会分散产卵,后代在土壤中产卵的时间也会延长。
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引用次数: 0
Macronutrient regulation in nymphs of the two-spotted cricket, Gryllus bimaculatus (Orthoptera: Gryllidae) 双斑蟋蟀(直翅目:蝼蛄)若虫的宏量营养素调节。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-27 DOI: 10.1016/j.jinsphys.2024.104684
Woomin Kwon , Kwang Pum Lee

Crickets have been extensively studied in recent insect nutritional research, but it remains largely unexplored how they balance the intake of multiple nutrients. Here, we used the nutritional geometry framework to examine the behavioural and physiological regulation of dietary protein and carbohydrate in nymphs of the two-spotted cricket, Gryllus bimaculatus (Orthoptera: Gryllidae). Growth, intake, utilization efficiencies, and body composition were measured from the eighth instar nymphs that received either food pairs or single foods with differing protein and carbohydrate content. When food choices were available, crickets preferentially selected a carbohydrate-biased protein:carbohydrate (P:C) ratio of 1:1.74. During this nutrient selection, carbohydrate intake was more tightly regulated than protein intake. When confined to nutritionally imbalanced foods, crickets adopted a nutrient balancing strategy that maximized the nutrient intake regardless of the nutrient imbalance, reflecting their omnivorous feeding habit. Intake was significantly reduced when crickets were confined to the most carbohydrate-biased food (P:C = 1:5). When nutrients were ingested in excess of the requirements, the post-ingestive utilization efficiencies of these nutrients were down-regulated, thereby buffering the impacts of nutrient imbalances on body nutrient composition. Crickets reared on the most carbohydrate-biased food (P:C = 1:5) suffered delayed development and reduced growth. Our data provide the most accurate description of nutrient regulation in G. bimaculatus and lay the foundation for further nutritional research in this omnivorous insect.

近年来的昆虫营养学研究对蟋蟀进行了广泛的研究,但对它们如何平衡多种营养物质的摄入量基本上仍未进行探讨。在这里,我们利用营养几何框架研究了双斑蟋蟀(直翅目:蟋蟀科)若虫对食物蛋白质和碳水化合物的行为和生理调节。我们测量了八龄若虫的生长、摄入量、利用效率和身体成分,这些若虫既可摄入成对的食物,也可摄入蛋白质和碳水化合物含量不同的单一食物。在有食物可供选择的情况下,蟋蟀优先选择碳水化合物比例为 1:1.74 的蛋白质:碳水化合物(P:C)。在这种营养选择过程中,碳水化合物的摄入量比蛋白质的摄入量受到更严格的控制。当被限制在营养失衡的食物中时,蟋蟀会采取一种营养平衡策略,无论营养失衡与否,都会最大限度地增加营养摄入量,这反映了蟋蟀的杂食习性。当蟋蟀只能摄入碳水化合物含量最高的食物(P:C = 1:5)时,摄入量会明显减少。当摄入的营养物质超过需要量时,这些营养物质的消化后利用效率会降低,从而缓冲营养失衡对体内营养成分的影响。使用碳水化合物偏重程度最高的食物(P:C = 1:5)饲养的蟋蟀发育迟缓,生长速度降低。我们的数据最准确地描述了G. bimaculatus的营养调节,为进一步研究这种杂食性昆虫的营养奠定了基础。
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引用次数: 0
Dynamic response of essential amino acid biosynthesis in Buchnera aphidicola to supplement sub-optimal host nutrition Buchnera aphidicola 中必需氨基酸生物合成对补充寄主营养不足的动态响应。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-27 DOI: 10.1016/j.jinsphys.2024.104683
Chloe C. Start, Catriona M.H. Anderson, Angharad M.R. Gatehouse, Martin G. Edwards

The endosymbiotic bacterium Buchnera aphidicola allows its host Acyrthosiphon pisum to utilise a nutritionally limited phloem sap diet without significant mortality by providing essential amino acids (EAAs), which it biosynthesises de novo via complex pathways consisting of multiple enzymes. Previous studies have reported how non-essential amino acids (NEAAs) provided by the host are utilised by B. aphidicola, along with how genes within the biosynthetic pathways respond to amino acid deficiency. Although the effect on B. aphidicola gene expression upon the removal of a single EAA and multiple NEAAs from the A. pisum diet has been reported, little is known about the effects of the complete simultaneous removal of multiple EAAs, especially branched-chain amino acids (BCAAs). To investigate this, A. pisum was provided with amino acid deficient diets ilv- (lacking isoleucine, leucine, valine) or thra- (lacking threonine, methionine, lysine). Due to their involvement in the production of several amino acids, the expression of genes ilvC, ilvD (both involved in isoleucine, leucine and valine biosynthesis) and thrA (involved in threonine, methionine and lysine biosynthesis) was analysed and the expression of trpC (involved in tryptophan biosynthesis) was used as a control. Survival was reduced significantly when A. pisum was reared on ilv- or thra- (P < 0.001 and P = 0.000 respectively) compared to optimal artificial diet and was significantly lower on ilv- (P < 0.001) than thra-. This is likely attributed to the EAAs absent from ilv- being required at higher concentrations for aphid growth, than those EAAs absent from thra-. Expression of ilvC and ilvD were upregulated 2.49- and 2.08-fold (respectively) and thrA expression increased 2.35- and 2.12-fold when A. pisum was reared on ilv- and thra- (respectively). The surprisingly large upregulation of thrA when reared on ilv- is likely due to threonine being an intermediate in isoleucine biosynthesis. Expression of trpC was not affected by rearing on either of the two amino acid deficient diets. To our knowledge this study has shown, for the first time, how genes within the biosynthetic pathways of an endosymbiont respond to the simultaneous complete omission of multiple EAAs as well as all three BCAAs (leucine, isoleucine, valine), from the host diet.

内共生细菌 Buchnera aphidicola 通过提供必需氨基酸(EAAs),使其宿主 Acyrthosiphon pisum 能够利用营养有限的韧皮部汁液而不会大量死亡。先前的研究报告了蚜虫如何利用宿主提供的非必需氨基酸(NEAAs),以及生物合成途径中的基因如何对氨基酸缺乏做出反应。尽管已经报道了从 A. pisum 的食物中移除单一 EAA 和多种 NEAAs 对 B. aphidicola 基因表达的影响,但对同时完全移除多种 EAA,尤其是支链氨基酸(BCAAs)的影响却知之甚少。为了探究这一问题,我们为 A. pisum 提供了氨基酸缺乏的日粮 ilv-(缺乏异亮氨酸、亮氨酸和缬氨酸)或 thra-(缺乏苏氨酸、蛋氨酸和赖氨酸)。由于它们参与多种氨基酸的生产,因此分析了基因 ilvC、ilvD(均参与异亮氨酸、亮氨酸和缬氨酸的生物合成)和 thrA(参与苏氨酸、蛋氨酸和赖氨酸的生物合成)的表达情况,并以基因 trpC(参与色氨酸的生物合成)的表达情况作为对照。与最适人工饲料相比,用 ilv- 或 thra- 饲养 A. pisum 的存活率明显降低(分别为 P < 0.001 和 P = 0.000),用 ilv- 饲养的存活率明显低于用 thra- 饲养的存活率(P < 0.001)。这可能是因为ilv-中缺乏的EAAs比thra-中缺乏的EAAs在蚜虫生长过程中需要更高浓度的EAAs。当 A. pisum 在 ilv- 和 thra- 上饲养时,ilvC 和 ilvD 的表达分别上调了 2.49 倍和 2.08 倍,thrA 的表达分别增加了 2.35 倍和 2.12 倍。在 ilv- 上饲养时,thrA 的表达出人意料地大幅上调,这可能是因为苏氨酸是异亮氨酸生物合成的中间体。使用两种氨基酸缺乏的日粮饲养都不会影响 trpC 的表达。据我们所知,这项研究首次显示了内共生体生物合成途径中的基因如何对宿主食物中同时完全缺乏多种 EAA 以及全部三种 BCAA(亮氨酸、异亮氨酸和缬氨酸)做出反应。
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引用次数: 0
Establishment of a rapid, cost-effective, and accurate method for assessing insect sperm viability 建立快速、经济、准确的昆虫精子活力评估方法。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-26 DOI: 10.1016/j.jinsphys.2024.104682
Mika Takeshima , Ayako Gotoh

High-quality sperm cells are crucial to reproductive success for both males and post-mating females in animals. Sperm viability, defined as the proportion of viable sperm cells, is used as a sperm quality index and this method has provided new insights into research on reproductive strategies. Sperm viability has been assessed by fluorescent staining of sperm cells. However, current staining protocols could potentially underestimate viability due to cell damage caused by cell treatments such as high dye concentration and long time for post-mounting. In this study, we established a method that enables rapid sperm viability assessment, has low sperm cell toxicity, and provides precise results regardless of operator expertise, and cost-effective using sperm cells from an ant, Crematogaster osakensis (Hymenoptera). First, to shorten the time for observation of a sufficient number of sperm cells, the volume per field of view was increased by height elevation between the glass slide and the coverslip, thereby we increased the number of sperm cells in a field of view. Second, to reduce sperm cell toxicity, we optimized the minimum dye concentration and incubation time using acridine orange (AO) and Hoechst in addition to SYBR 14 and propidium iodide (PI), which has been used in most previous studies. We determined the optimal protocol to be 1 µg/mL AO and 150 µM PI without incubation. Besides, we automated counting sperm cells with ImageJ software and combined with manual correction for more accurate results. We employed the improved method for sperm samples from mealworm beetles (Tenebrio molitor) and silkmoths (Bombyx mori). This method, established through our study, will advance research on reproductive strategies, including sperm competition and sperm quality maintenance in females.

高质量的精子细胞对雄性动物和交配后的雌性动物的繁殖成功至关重要。精子存活率(定义为存活精子细胞的比例)被用作精子质量指标,这种方法为生殖策略研究提供了新的视角。然而,目前的染色方案可能会低估精子的存活率,原因是高浓度染料和长时间贴装等细胞处理方法会造成细胞损伤。在这项研究中,我们利用蚂蚁(膜翅目)Crematogaster osakensis的精子细胞,建立了一种能快速评估精子活力、精子细胞毒性低、结果精确且不受操作者专业知识影响的方法。首先,为了缩短观察足够数量精子细胞的时间,我们通过提高玻璃载玻片和盖玻片之间的高度来增加每个视野的体积,从而增加了视野中精子细胞的数量。其次,为了降低精子细胞的毒性,我们优化了最小染料浓度和孵育时间,除了使用 SYBR14 和碘化丙啶(PI)之外,还使用了吖啶橙(AO)和 Hoechst。我们确定最佳方案为 1 µg/mL AO 和 150 µM PI,无需孵育。此外,我们还利用 ImageJ 软件对精子细胞进行了自动计数,并结合人工校正以获得更准确的结果。我们对黄粉虫(Tenebrio molitor)和丝蛾(Bombyx mori)的精子样本采用了改进的方法。通过我们的研究建立的这一方法将推动生殖策略的研究,包括精子竞争和雌性精子质量的维持。
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引用次数: 0
Long-term impacts of egg quiescence and Wolbachia infection on lipid profiles in Aedes aegypti: Ovarian roles in lipid synthesis during reproduction 卵静止和沃尔巴克氏体感染对埃及伊蚊脂质特征的长期影响:卵巢在繁殖期间脂质合成中的作用。
IF 2.3 2区 农林科学 Q1 ENTOMOLOGY Pub Date : 2024-07-10 DOI: 10.1016/j.jinsphys.2024.104674

Wolbachia, an endosymbiotic bacterium, relies on nutrients from its host to complete its life cycle. The presence of Wolbachia strain wAlbB in the mosquito Aedes aegypti during egg or larval stages affects the host’s development, leading to the absence of developed and visible ovaries in adult mosquito females. In this study, we investigated the impacts of egg quiescence and Wolbachia infection on lipid profiles of adult Ae. aegypti females, and discerned the role of ovaries in lipid synthesis in the reproductive process. The lipidomes of Wolbachia infected and uninfected female individuals at various developmental stages were quantitatively analyzed by LC-MS/MS. Lipidomic change patterns were systematically further investigated in wAlbB-infected fertile females and infertile females following blood feeding. Prolonged egg quiescence induced a shortage of acyl-carnitine (CAR) and potentially impacted some molecules of diacyl-phospholipid (diacyl-PL) and sphingolipid (SL) in young adult mosquitoes. After the first gonotrophic cycle, infertile females accumulated more CAR and lyso-phospholipid (lyso-PL) than fertile females. Then in the second gonotrophic cycle, the patterns of different lipid groups remained similar between fertile and infertile females. Only a small proportion of molecules of triglyceride (TG), phospholipid (lyso-PL and diacyl-PL) and ceramide (Cer) increased exclusively in fertile females from 0 h to 16 h post blood meal, suggesting that the generation or prescence of these lipids rely on ovaries. In addition, we found cardiolipins (CL) might be impacted by Wolbachia infection at the egg stage, and infected mosquitoes also showed distinct patterns between fertile and infertile females at their second gonotrophic cycle. Our study provides new insights into the long-term influence of Wolbachia on lipid profiles throughout various life stages of mosquitoes. Additionally, it suggests a role played by ovaries in lipid synthesis during mosquito reproduction.

沃尔巴克氏菌是一种内共生细菌,依靠宿主提供的营养来完成其生命周期。在埃及伊蚊的卵或幼虫阶段,沃尔巴克氏菌株 wAlbB 的存在会影响宿主的发育,导致成年雌蚊没有发育和可见的卵巢。在这项研究中,我们研究了卵静止和沃尔巴克氏体感染对成年埃及伊蚊雌虫脂质图谱的影响,并发现了卵巢在生殖过程中脂质合成的作用。采用 LC-MS/MS 对感染了沃尔巴克氏菌和未感染沃尔巴克氏菌的雌性个体在不同发育阶段的脂质体进行了定量分析。进一步系统地研究了经 wAlbB 感染的可育雌性个体和经血饲喂后的不育雌性个体的脂质体变化规律。长时间的卵静止诱导了酰基肉碱(CAR)的短缺,并可能影响年轻成蚊体内的一些二酰磷脂(diacyl-PL)和鞘脂(SL)分子。在第一个生殖周期后,未育雌蚊比已育雌蚊积累了更多的 CAR 和溶血磷脂(溶血磷脂)。然后,在第二个生殖周期中,可育雌蚊和不可育雌蚊的不同脂质组的模式仍然相似。只有一小部分甘油三酯(TG)、磷脂(溶菌酶-磷脂和二酰基-磷脂)和神经酰胺(Cer)分子在血餐后0小时至16小时内只在可育雌鼠体内增加,这表明这些脂质的产生或存在依赖于卵巢。此外,我们发现心磷脂(CL)可能会在卵阶段受到沃尔巴克氏体感染的影响,受感染的蚊子在其第二个生殖腺周期也表现出可育雌蚊和不育雌蚊的不同模式。我们的研究为了解沃尔巴克氏菌对蚊子各生命阶段脂质特征的长期影响提供了新的视角。此外,它还表明卵巢在蚊子繁殖过程中的脂质合成中发挥了作用。
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引用次数: 0
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Journal of insect physiology
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